Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase.

摘要: Nitric oxide, which accounts for the biological activity of endothelium-derived relaxing factor (EDRF), is synthesized in endothelial cells from L-arginine by nitric oxide synthase (NOS). We report here cloning and functional expression a cDNA encoding human NOS. Oligonucleotides corresponding to amino acid sequences shared cytochrome P450 reductase recently identified brain NOS were amplify partial bovine cell NOS-related protein. This was used isolate vascular The translated protein 1294 acids long shares 52% its sequence with Using RNA blot hybridization, abundant mRNA detected unstimulated umbilical vein cells. To determine protein, we ligated into an vector transfected it NIH3T3 Cells expressing this contained NADPH diaphorase activity, histochemical marker In co-culture assays, production increased guanylate cyclase reporter rat fetal lung fibroblasts. addition, NOS-catalyzed conversion arginine citrulline significantly A23187, calcium ionophore. Isolation calcium-regulated, constitutively expressed NOS, capable producing EDRF blood vessels, will accelerate characterization role enzyme normal abnormal regulation tone.